Assessment of Risks and Benefits of Using Antibiotics Resistance Genes in Mesenchymal Stem Cell-Based Ex-Vivo Therapy

Recently, ex-vivo gene therapy has emerged as a promising approach to enhance the therapeutic potential of mesenchymal stem cells (MSCs) by introducing functional genes in vitro. Here, we explored the need of using selection markers to increase the gene delivery efficiency and evaluated the potential risks associated with their use in the manufacturing process. We used MSCs/CD that carry the cytosine deaminase gene (CD) as a therapeutic gene and a puromycin resistance gene (PuroR) as a selection marker. We evaluated the correlation between the therapeutic efficacy and the purity of therapeutic MSCs/CD by examining their anti-cancer effect on co-cultured U87/GFP cells. To simulate in vivo horizontal transfer of the PuroR gene in vivo, we generated a puromycin-resistant

Preclinical Study on Biodistribution of Mesenchymal Stem Cells after Local Transplantation into the Brain

Therapeutic efficacy of mesenchymal stem cells (MSCs) is determined by biodistribution and engraftment in vivo.Compared to intravenous infusion, biodistribution of locally transplanted MSCs are partially understood. Here, we performed a pharmacokinetics (PK) study of MSCs after local transplantation. We grafted human MSCs into the brains of immune-compromised nude mice. Then we extracted genomic DNA from brains, lungs, and livers after transplantation over a month. Using quantitative polymerase chain reaction with human Alu-specific primers, we analyzed biodistribution of the transplanted cells. To evaluate the role of residual immune response in the brain, MSCs expressing a cytosine deaminase (MSCs/CD) were used to ablate resident immune cells at the injection site. The majority of the Alu signals mostly remained at the injection site and decreased over a week, finally becoming undetectable after one month. Negligible signals were transiently detected in the lung and liver during the first week. Suppression of Iba1-positive microglia in the vicinity of the injection site using MSCs/CD prolonged the presence of the Alu signals.After local transplantation in xenograft animal models, human MSCs remain predominantly near the injection site for limited time without disseminating to other organs. Transplantation of human MSCs can locally elicit an immune response in immune compromised animals, and suppressing resident immune cells can prolong the presence of transplanted cells. Our study provides valuable insights into the in vivo fate of locally transplanted stem cells and a local delivery is effective to achieve desired dosages for neurological diseases.

Factors Influencing Self-Rated Oral Health in Elderly People Residing in the Community: Results from the Korea Community Health Survey, 2016

ObjectivesThe aim of this study was to examine the factors influencing perceived oral health in elderly individuals residing in the community.MethodsThis study used raw data from the Korea community health survey, 2016. Of the 64,223 participants that were elderly (aged ≥ 65 years), 61,280 (95.4%) were included for analysis. Self-rated oral health was the dependent variable and 6 independent variables including age, gender, type of area of residence (metropolitan or provincial), educational level, income, and living status with spouse were assessed. Oral function was studied based on mastication, pronunciation, and use of dentures, and oral health behavior included brushing teeth after breakfast, after lunch, after dinner, and before sleep). The EQ-5D questionnaire measured health-related quality of life (mobility, self-care, usual activities, pain/discomfort and anxiety/depression).ResultsAmong the general characteristics, age, gender, educational level, income, and living status with spouse were the factors that affected self-rated oral health. Mastication, pronunciation, use of dentures, and brushing after lunch, dinner, and before sleep were the factors that influenced self-rated oral function. All domains of the EQ-5D (pain/discomfort, mobility, self-care, usual activities, and anxiety/depression) were factors that affected self-rated oral health.ConclusionThe results of the current investigation suggest that the development of management and education strategies for oral health promotion in the elderly, should focus on improving oral function and oral health behavior, taking into account the socio-economic and demographic characteristics that have been shown to be associated with poor self-rated oral function.

Retrovirus-mediated transduction of a cytosine deaminase gene preserves the stemness of mesenchymal stem cells

Human mesenchymal stem cells (MSCs) have emerged as attractive cellular vehicles to deliver therapeutic genes for ex-vivo therapy of diverse diseases; this is, in part, because they have the capability to migrate into tumor or lesion sites. Previously, we showed that MSCs could be utilized to deliver a bacterial cytosine deaminase (CD) suicide gene to brain tumors. Here we assessed whether transduction with a retroviral vector encoding CD gene altered the stem cell property of MSCs. MSCs were transduced at passage 1 and cultivated up to passage 11. We found that proliferation and differentiation potentials, chromosomal stability and surface antigenicity of MSCs were not altered by retroviral transduction. The results indicate that retroviral vectors can be safely utilized for delivery of suicide genes to MSCs for ex-vivo therapy. We also found that a single retroviral transduction was sufficient for sustainable expression up to passage 10. The persistent expression of the transduced gene indicates that transduced MSCs provide a tractable and manageable approach for potential use in allogeneic transplantation.

Clinical Effects of Oseltamivir in Children with Influenza in Busan, in the First Half of 2004 / 소아과

PURPOSE: Although influenza is one of the most important causes of acute respiratory tract infections in children, effective antiviral therapies are not common and there are only a few clinical studies on treatment of influenza in children. We evaluated the efficacy of oseltamivir in the treatment of naturally aquired influenza in children during the first half of 2004 in Busan. METHODS: From January 2004 to June 2004, throat swabs and nasal washes were performed and cultured for the isolation of influenza virus and tested by rapid antigen detection test (QuickVue influenza test) in children with suspected influenza infections. The children who responded positively to the QuickVue influenza test, we divided into two groups: an oseltamivir treatment group and a control group. We compared their clinical symptoms (including fever duration) and diagnosis. The medical records of patients with influenza virus infection were reviewed retrospectively. RESULTS: A total of 621 individuals were suspected of influenza infection. Influenza viruses were isolated in 79 (17.2 percent) out of 621 patients examined. QuickVue influenza tests were positive in 181 cases. The treatment group (83 individuals) received oseltamivir twice daily for 5 days, and the control group (99 individuals) were administered only symptom relief medicine. There was no differences between the two groups in clinical diagnosis and symptoms. Oseltamivir treatment reduced the fever duration and other respiratory symptoms. There were no adverse events associated with oseltamivir treatment. CONCLUSION: Our data suggest that oral oseltamivir treatment reduces the fever duration and other respiratory symptoms of acute influenza without side effects in children.

A Case of Severe Neonatal Hypertension in Twin-Twin Transfusion Syndrome

Twin-twin transfusion syndrome is attributed to an unbalanced blood flow between the donor and the recipient twin, but the exact pathophysiology remains incompletely understood. Despite active prenatal management, fetal twin-twin transfusion syndrome is still associated with a substantial residual perinatal mortality and morbidity. The donor twin progressively becomes anemic, growth restricted, oliguric and has oligohydramnios, where as the recipient becomes plethoric, polyuric and has polyhydramnios and in the most severe cases develops cardiomegaly, congestive heart failure and hypertension. We report a case of severe systemic hypertension developed in recipient twin.

Spontaneous Ileal Perforation in Very Low Birth Weight Infant without Evidence of Necrotizing Enterocolitis

Ongoing advances in the treatment of very-low-birth-weight infants have confronted us with a growing number of infants susceptible to acquired illnesses of the gastrointestinal tract. Although necrotizing enterocolitis has been regarded as the major cause of gastrointestinal perforation in preterm infants, the incidence of spontaneous perforation occurred in an apparently normal bowel is increasing. The risk factors for spontaneous intestinal perforation include prematurity, twin pregnancies, perinatal asphyxia, prior use of umbilical artery catheter, use of indomethacin and/or steroid, and bacterial or fungal sepsis. We report a case of spontaneous ileal perforation occurred in very low birth weight infant who was successfully treated with emergency operation.

Successful Long Term Antifungal Agent Therapy for Renal Candidiasis without Surgical Drainage in a Very Low Birth Weight Infant

Improved survival rate of premature infants requiring intensive care lead into an increased risk for nosocomial infections such as disseminated fungal infection. Neonatal candida sepsis has become one of the most important causes of neonatal morbidity and mortality. The most common site of end organ involvement in premature infants with candidemia is the kidney. But no consensus has been reached concerning the treatment of candidemia in the newborn. We recently experienced a case of premature infant who was diagnosed as renal candidiasis with microabscess formation due to Candida Albicans and patient was treated successfully with long term liposomal amphotericin B and fluconazole therapy without surgical drainage.

A Case of Diabetic Nephropathy without Microalbuminuria in Type 1 Diabetes / 대한소아내분비학회지

Diabetes is a rapidly increasing heath care problem all over the world due to increased prevalence during past decade. Diabetic nephropathy develops in 25-30% of patients with type 1 diabetes and is the leading cause of end stage renal disease. Diabetic nephropathy is characterized by persistent proteinuria, decline in renal function, hypertension and increased cardiovascular morbidity and mortality. Early detection of diabetic nephropathy risk is an important goal because early diagnosis and treatment prevent advanced renal damage and other diabetic complications. Increased urinary albumin excretion rate is widely accepted as the first clinical sign of diabetic nephropathy. However, reduced glomerular filtration or hypertension could be the first manifestation in some diabetic patients. We need improved markers and predictors of diabetic nephropathy risk. We report a case of diabetic nephropathy and decreased glomerular filtration rate (GFR) without microalbuminuria occcured in type 1 diabetic patient.

A Clinical Study of Aseptic Meningitis in the Busan Area in 2002

PURPOSE: There was a outbreak of aseptic meningitis in Busan, 2002. We report the clinical features and causative viruses. METHODS: Two hundred seventy six children with aseptic meningitis who were admitted to the Department of Pediatrics, Maryknoll Hospital between January and December, 2002 were included. CSF, stool and throat swab viral cultures were done in 244 of these children. RESULTS: The male to female ratio was 1.4 : 1. Age of patients varied from five months to fourteen years old. Average age was 5.7+/-6.2 years old and mostly between one and four years.(41.3%) Monthly distribution revealed that the number of patients increased from May to July. The main symptoms were fever, headache and vomiting in this order. Respiratory tract infection symptom was associated from June to July and headache and nausea without fever were characteristically observed in children more than 10 years old from November to December. In peripheral blood examination, leukocytosis(WBC>10,000/mm3) showed in 34.8%, ESR was increased in 56.1%, and CRP was positive value in 61%. Therefore differential diagnosis was difficult through peripheral blood examination. CSF findings revealed mean leukocyte count 86.5+/-180.2/mm3, protein 41.7+/-32.9 mg/dL, glucose 56.4+/-9.9 mg/dL. Median hospitalized period was 4.7+/-7.2 days and compared with non-tapping group, hospitalized period was shorter and subsidance of symptoms was faster, therefore antibiotics injection period was shorter in the spinal tapping group. Virus was isolated in 31 cases of 244. The causative agents were echovirus 6, echovirus 9, echovirus 25, coxsakie virus B3, B4. CONCLUSION: There was an epidemics of aseptic meningitis in Busan, 2002; the causative agent was echovirus 6, 9, 25, coxsakie virus B3, B4.

Evaluation of Timing of Voiding Cystourethrogram after Urinary Tract Infection

PURPOSE: Urinary tract infection is a common problem in children. To evaluate for reflux most authorities recommend a voiding cystourethrogram 3 to 6 weeks after the first urinary tract infection. But during the 3 to 6 weeks interval, patients may fail to show up for the scheduled VCUG and thus risk for loss of follow up. We analyzed patient's records to evaluate whether the timing of VCUG after UTI influenced the prevalence or severity of VUR. METHODS: We retrospectively reviewed 213 children diagnosed with UTI from March 1997 to December 2000. These children were divided into 2 groups according to whether they had VCUG scheduled to be performed either within 1 weeks after the diagnosis of UTI (Group A) or later than 1 week after the diagnosis(Group B). We compared the presence and severity of reflux in the 2 groups. RESULTS: Reflux was present in 19% of the patients studied within 1 week after UTI and in 18% of those studied after 1 week. This difference was not statistically significant. Whereas 100% of the scheduled VCUGs in the Group A were performed, only 48% of those scheduled in the Group B were performed. This difference is statistically significant. CONCLUSION: Because there was no significant difference between the presence or severity of reflux and timing of VCUG after UTI, we suggest that a hospitalized patient with UTI should have VCUG performed before discharge.

Comparison of the Effectiveness of Phototherapy for Nonhemolytic Hyperbilirubinemia in Breast-fed and Formula-fed Infants

PURPOSE: This study was designed to evaluate the efficacy of phototherapy for nonhemolytic hyperbilirubinemia in breast-fed and formula-fed infants and infants receiving formula and breast milk. METHODS: Full-term healthy infants with nonhemolytic hyperbilirubinemia (bilirubin concentration > or =13.0 mg/dL) were treated with conventional phototherapy by using daylight fluorescent lamps. Three groups of infants were studied : group A, formula-fed infants; group B, breast-fed infants; group C, infants receiving formula and breast milk. All patterns of feeding started at birth. Phototherapy was terminated only when bilirubin concentration had decreased to less than 10.0 mg/dL. RESULTS: A total of 163 infants were studied. Phototherapy was highly effective in reducing the bilirubin concentration in all three groups, however, its efficacy of in group B was poorer compared with the other two groups. The duration of photopherapy and weight loss during phototherapy were not significantly different in the three groups. Decrease rates of bilirubin concentration during phototherapy in group A and C were significantly faster than that of group B. CONCLUSION: The response to phototherapy of group B infants was significantly slower than those of group A and C infants. The addition of formula to the feedings for totally breast-fed infants, without suspension of breast feeding, would enhance the efficacy of phototherapy and reduce exposure time.

A Case of Extrapelvic Endometriosis with Abdominal Pain in a 13-Year-Old Girl

Endometriosis is defined as the presence of endometrial tissue, e.g. gland and stroma, outside the uterus. Extrapelvic endometriosis is a rare condition but it occurrs in the gastrointestinal tract, kidney, pancreas, umbilicus, perianal area, abdominal scar tissue, lung or central nervous system except spleen. We have experienced a case of extrapelvic endometriosis in an adolescent girl, who complained of severe left upper quadrant abdominal pain and vomiting. The lesion was diagnosed by abdominal ultrasonography and computerized tomography, and confirmed histologically by exploratory laparotomy and mass excision biopsy. During menstration after surgery, she had no complaint of abdominal pain nor dysmenorrhea. We report this case with a review of associated literature.

A Case of Parry-Romberg Syndrome in Neonate

Parry-Romberg syndrome(Progressive hemifacial atrophy), described in the last century by Parry(1825) and Romberg(1846), is a very rare disorder characterized by a slowly progressive and self-limited unilateral(rarely bilateral) atrophy of the faces affecting variably the skin, subcutaneous fat tissues, musculature, connective tissue, cartilage and bones. And this disorder is usually accompanied by contralateral Jacksonian epilepsy, trigerminal neuralgia, and changes in the eyes and hair. The onset is slow and progressive, starting at 5-15 years of age and lasting from 2-10 years, ending with the face being "burned out". There are a few cases of this disease which presented during the neonatal period. This disorder seems to affect females more than males, and its etiology and incidence has yet to be determined. Trauma, infection with a slow virus, sympathetic dysfunction, immunological abnormality and cranial vascular malformation are proposed causes. No typical or consistent neuropathologic findings occur. No specific treatment for the syndrome exists; however, various reconstructive surgical procedures can have in reasonably good cosmetic effects, as well as antiinflammatory or immunosuppressive treatment. We report a case of Parry-Romberg syndrome, which was presented at 1 month of age, and has progressd to contralateral hemiparesis.

A Case of Primary Gastric Lymphoma in Puberty / 대한소아소화기영양학회지

Primary gastric lymphoma is relatively uncommon, accounting for 3% to 8% of all malignancies arising in the stomach. The most common symptom is abdominal pain, closely followed by weight loss, anorexia, weakness due to anemia, nausea, and vomiting. The diagnosis of gastric lymphoma usually requires a biopsy at the time of gastroscopy or laparotomy. Microscopically, the vast majority of gastric lymphoid tumors are non-Hodgkin's lymphomas of B cell origin. Survival rates for all types of gastric lymphoma generally exceed those for adenocarcinoma and other malignancies of the stomach. We experienced one case of primary gastric lymphoma in puberty with a brief review of the literature.

Basophil Histamine Releasability in Asthmatic Children

PURPOSE: The release of histamine from human basophils is controlled by an intrinsic, as yet unidentified, cellular property termed "releasability." We carried out this study to ascertain whether there was any difference in the releasability of basophils from asthmatic children compared to those from normal children. We intended also to clarify the correlation between the releasability and the atopic status of asthma. METHODS: We selected nineteen atopic asthmatic, eighteen nonatopic asthmatic and fourteen normal children for this study. Suspensions of leukocytes were isolated and stimulated with calcium ionophore A23187, anti-IgE and D. pteronyssinus antigen. After incubation, the supernatant was assayed for histamine with an automated fluorometric technique. RESULTS: Basophil histamine release with anti-IgE was different in three groups. Anti-IgE caused significantly more basophil histamine release in asthmatic children than in nomal children. Atopic asthmatic group showed greater basophil histamine release with anti-IgE than nonatopic asthmatic group. D. pteronyssinus antigen caused the significant amount of histamine release only in atopic asthmatic group. CONCLUSIONS: Our data suggests that basophils from asthmatic children are characterized by a specific increase in IgE mediated histamine releasability. The difference of histamine releasability with anti-IgE between atopic and nonatopic asthmatic children may be due to the heterogeneity of IgE bound to cell surface, or may be due to the degree of the basophil activation by cytokines such as IL3. The specific release of histamine with D. pteronyssinus antigen in atopic asthmatic group suggests that the basophil histamine release test can be used to diagnose the causing antigen.

A Case of Congenital Factor VII Deficiency Associated with Intraventricular Hemorrhage and Hydrocephalus

Congenital factor Vll deficiency is a rare bleeding disorder with an estimated incidence of 1 in 500,000. It is inherited as an autosomal recessive pattern with variable expression and high penetrance. In severely affected patients, repeated hemarthroses, chronic crippling hemarthropathy, and dangerous hematomas can occur. Other types of hemorrhage include epistaxis, menorrhagia, hematuria, gastrointestinal and gingival bleeding. Fetal cerebral hemorrhage has been reported, although less frequently than in severe hemophilia A or B. It is characterized by normal partial thromboplastin time and prolonged prothrombin time. Definitive diagnosis rests on a specific assay for factor Vll clotting activity. Replacement therapy is necessary to control the hemorrhage. Conventional prophylaxis and therapy in this disorder have consisted of fresh frozen plasma (FFP) or prothrombin complex concentrate. We experienced a case of intraventricular hemorrhage and hydrocephalus in a 4-year-old girl who had been diagnosed with congenital factor Vll deficiency during her neonatal period. She presented with episodes of frontal headache, frequent vomiting and malnutrition. We report this case with a brief review and related literatures.

Studies on familial of basophil histamine releasability / 천식및알레르기

BACKGROUND: Bronchial asthma is a complex genetic disorder. Although serum IgE level and bronchial hyperresponsiveness are well known to be under genetic control, the influence of genetic factors on basophil releasability has been seldom studied. OBJECTIVE: The present study was carried out to investigate whether genetic factors may influence the basophil histamine releasability. MATERIALS AND METHODS: We studied 50 children, 32 with atopic asthma (AA) and 18 normal control (NC), and their parents. Suspensions of leukocytes were isolated and stimulated with Ca ionophore and anti-IgE antibody. Then, histamine in the supernatant was as-sayed by an automated fluorometric analyzer. RESULTS: Among the probands, AA children had a significantly higher anti-IgE induced histamine release than NC children. In contrast, Ca ionophore-induced histamine release was similar between the two groups. Ca ionophore-induced or anti-IgE-induced histamine release was not significantly different between parents of AA children and those of NC children. However, the maximal histamine release by Ca ionophore in parents had a significant correlation with that of probands, whereas the values by anti-IgE were not correlated between probands and their parents. CONCLUSION: We confirmed that basophils from patients with atopic asthma are characterized by a specific increase in IgE-mediated histamine release. The significant correlation of Ca ionophore-induced maximal histamine release between children and their parents suggests that genetic factors may play an important role in the control of non-IgE-mediated relessability from basophils.

Tthe influences of parental atopy and bronchial hyperresponsiveness on the development of asthma agter early childhood wheezing / 천식및알레르기

BACKGROUND: Many young children suffer from wheezing illness during viral respiratory infection, and some of them experience wheezing many years later and ultimately develop bronchial asthma. It is not clear whether atopy or bronchial hyperresponsiveness in the family is a significant risk factor for asthma in this clinical setting. Objective : To examine the genetic basis for the development of asthma after early childhood wheezing. Materials and METHODS: A measurement of serum total IgE concentration, skin prick test to common inhalant allergens, and methacholine bronchial provocation test were performed in 29 asthmatic children and their parents, and 22 non-asthmatic children with the past history of wheezing illness during the first three years of age and their parents. A questionnaire was performed to assess the presence of asthma and allergic rhinitis in the parents. RESULTS: Positive skin test response to common inhalant allergens was more prevalent in asthmatics than in non-asthmatics(67.8% vs. 27.2%). Serum total IgE concentration was significantly higher in asthmatics than in non-asthmatics(geometric mean: 173 vs. 83 IU/ ml). Positive skin test response to comman inhalant allergens was more prevalent in parents of asthmatics than in thoae of non-asthmatics(51.7% vs. 25.0%), but serum total IgE level was not different between the two groups(geometric mean: 132 vs. 120 IU/ml). Positive rate of methacholine bronchial provocation test, geometric mean of PC20-methacholine, and BR index were not different between the parents of asthmatics and non-asthmatics (18.1% vs. 13.9%; 164 vs. 180 mg/ml; 1.154+-0.077 vs. 1.055+-0.068, respectively). CONCLUSION: It is suggested that personal atopy is important in the development of asthma after early childhood wheezing, and parental atopy rather than bronchial hyperresponsiveness is a risk factor for the development of childhood asthma in this clinical setting.

Immunohistochemical Study of Bcl-2 Oncoprotein Expression in Childhood Non-Hodgkin's Lymphoma

PURPOSE: The aim of the present study was to assess the clinicopatholgical significance of Bcl-2 oncoprotein expression in childhood non-Hodgkin's lymphoma (NHL). METHODS: We have assessed 16 cases of childhood NHL during last 6 years from 1990 to 1995. Bcl-2 oncoprotein expression has been semiquantitatively analyzed in paraffin sections from 16 cases of childhood NHL with 39 control cases of adult NHL. The expression of Bcl-2 oncoprotein was correlated with histologic grade, immunophenotype, proliferative activity as measured by immunostain of Ki-67 antigen, clinical stage, and survival rate (event free survival rate, EFS). RESULTS: 1) Bcl-2 oncoprotein expression significantly decreased according to increase of histological grade (P0.05). 5) EFS of patients was significantly correlated with Bcl-2 oncoprotein expression, that is, reduced EFS was demonstrated in the patients with low Bcl-2 oncoprotein expression (P<0.05). CONCLUSIONS: Thus Bcl-2 oncoprotein, as demonstrated immunohistochemically in routinely paraffin embedded tissue, can be restrictively used in prediction of prognosis and grade of childhood NHL, in aggrement with the role of Bcl-2 oncoprotein in blocking of apoptosis and indirect contribution to increase of cellular proliferative activity of NHL.